Purification and Properties of Alcohol Dehydrogenase from the Acid-and Enthanol-Tolerant Yeast Candida solicola

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Two alcohol dehydrogenases(EC1.1.1.1)from tha acid- and ethanol-tolerant yeast Candida solicola WY-1 have been purified and characterized. The microbial strain cultured in a medium containing ethanol as a sole carbon source was disrupted in a Dyno-mill. From the cell-free extract obtained by centrifugation at 105,000×g for 60min, two alcohol dehydrogenases(ADH 1,ADH 2)were separated by DEAE-Toyopearl 650M chromatography. ADH 1 was further purified by affinity chromatography using Matrex Blue A, ADH 2 was purified by chromatofocusing using Polybuffer Exchanger(PBE)94 and affinity chromatography using Matrex Blue A. ADH 1 and ADH 2 had the same optimum pH, 7.0. ADH 1 was stable between pH7.0 and 7.5,and ADH 2 at pH7.0. The molecular weights of ADH 1 and ADH 2 were calculated to be about 160,000 and 162,000,while the isoelectric points were 5.3 and 5.25,respectively. The optimum temperature of ADH 1 was 30℃, while that of ADH 2 was 55℃. ADH 1 was stable at temperatures below 50℃, whereas ADH 2 was unstable at temperatures above 25℃.
社団法人日本生物工学会の論文
1991-05-25